Scanning electron microscope assessment of exocytotic changes in purified gonadotropes

Abstract These studies were undertaken to characterize the exocytotic changes in purified gonadotropes by three-dimensional imaging using scanning electron microscopy. Rat gonadotropes were purified using a fluorescence-activated cell sorter and an argon laser treatment system. The purified gonadotropes were stimulated with GnRH under various conditions and fixed for scanning electron microscopy. After the GnRH stimulation, many 'hole' structures (diameter 0·1–0·5 μm) were observed on the cell surface, and notably the population of cells with 10 or more holes was clearly increased. The pattern of the time-course of the changes in this population was perfectly consistent with the LH secretory profile of pituitary cells, and their formation of the cells with 10 or more holes was completely inhibited by pretreatment with a GnRH antagonist. Our data suggest that the hole structure represents an exocytotic opening site and that regulated exocytosis in purified gonadotropes can be evaluated by scanning electron microscopy. This method may be widely applicable to other endocrine cells. Journal of Endocrinology (1995) 144, 193–200

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Scanning and Transmission Microscopy of Nematocyst Batteries in Epitheliomuscular Cells of Hydra

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066097 ◽

1971 ◽

Vol 29 ◽

pp. 410-411 ◽

Cited By ~ 1

Author(s):

Jane A. Westfall ◽

S. Yamataka ◽

Paul D. Enos

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Osmium Tetroxide ◽

External Surface ◽

Three Dimensional ◽

Surface Structures ◽

Transmission Microscopy ◽

Transmission Electron ◽

Freeze Dried ◽

Scanning Electron

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).

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Scanning Electron Microscopy and Electron Microprobe Analysis of Malignant Cell Cultures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100561 ◽

1968 ◽

Vol 26 ◽

pp. 164-165

Author(s):

R. I. Johnsson-Hegyeli ◽

A. F. Hegyeli ◽

D. K. Landstrom ◽

W. C. Lane

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Cell Cultures ◽

Electron Microprobe ◽

Microprobe Analysis ◽

Electron Microprobe Analysis ◽

Three Dimensional ◽

Malignant Cell ◽

Interference Microscopy ◽

Scanning Electron

Last year we reported on the use of reflected light interference microscopy (RLIM) for the direct color photography of the surfaces of living normal and malignant cell cultures without the use of replicas, fixatives, or stains. The surface topography of living cells was found to follow underlying cellular structures such as nuceloli, nuclear membranes, and cytoplasmic organelles, making possible the study of their three-dimensional relationships in time. The technique makes possible the direct examination of cells grown on opaque as well as transparent surfaces. The successful in situ electron microprobe analysis of the elemental composition and distribution within single tissue culture cells was also reported.This paper deals with the parallel and combined use of scanning electron microscopy (SEM) and the two previous techniques in a study of living and fixed cancer cells. All three studies can be carried out consecutively on the same experimental specimens without disturbing the cells or their structural relationships to each other and the surface on which they are grown. KB carcinoma cells were grown on glass coverslips in closed Leighto tubes as previously described. The cultures were photographed alive by means of RLIM, then fixed with a fixative modified from Sabatini, et al (1963).

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The time course of calcium deposition in shells of the barnacle (Balanus amphititre amphititre) during cyprid-juvenile metamorphosis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100168621 ◽

1994 ◽

Vol 52 ◽

pp. 178-179

Author(s):

Nancy R. Wallace ◽

Craig C. Freudenrich ◽

Karl Wilbur ◽

Peter Ingram ◽

Ann LeFurgey

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Time Course ◽

Vertical Plane ◽

Mantle Cavity ◽

Qualitative Assessment ◽

Calcium Deposition ◽

Free Swimming ◽

Freeze Dried ◽

Scanning Electron

The morphology of balanomorph barnacles during metamorphosis from the cyprid larval stage to the juvenile has been examined by light microscopy and scanning electron microscopy (SEM). The free-swimming cyprid attaches to a substrate, rotates 90° in the vertical plane, molts, and assumes the adult shape. The resulting metamorph is clad in soft cuticle and has an adult-like appearance with a mantle cavity, thorax with cirri, and incipient shell plates. At some time during the development from cyprid to juvenile, the barnacle begins to mineralize its shell, but it is not known whether calcification occurs before, during, or after ecdysis. To examine this issue, electron probe x-ray microanalysis (EPXMA) was used to detect calcium in cyprids and juveniles at various times during metamorphosis.Laboratory-raised, free-swimming cyprid larvae were allowed to settle on plastic coverslips in culture dishes of seawater. The cyprids were observed with a dissecting microscope, cryopreserved in liquid nitrogen-cooled liquid propane at various times (0-24 h) during metamorphosis, freeze dried, rotary carbon-coated, and examined with scanning electron microscopy (SEM). EPXMA dot maps were obtained in parallel for qualitative assessment of calcium and other elements in the carapace, wall, and opercular plates.

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Faculty Opinions recommendation of Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1022557.793507804 ◽

2015 ◽

Author(s):

Alpha Yap

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Three Dimensional ◽

Face Scanning ◽

Block Face ◽

Scanning Electron

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Three-dimensional relationships between tumor cells and microcirculation with double cyanine immunolabeling, laser scanning confocal microscopy, and computer-assisted reconstruction: an alternative to cast corrosion preparations.

Journal of Histochemistry & Cytochemistry ◽

10.1177/42.5.7908912 ◽

1994 ◽

Vol 42 (5) ◽

pp. 681-686 ◽

Cited By ~ 27

Author(s):

V Rummelt ◽

L M Gardner ◽

R Folberg ◽

S Beck ◽

B Knosp ◽

...

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Blood Vessels ◽

Tumor Cells ◽

Laser Scanning ◽

Three Dimensional ◽

Melanoma Cells ◽

Tumor Blood Vessels ◽

The Relationship ◽

Scanning Electron

The morphology of the microcirculation of uveal melanomas is a reliable market of tumor progression. Scanning electron microscopy of cast corrosion preparations can generate three-dimensional views of these vascular patterns, but this technique sacrifices the tumor parenchyma. Formalin-fixed wet tissue sections 100-150 microns thick from uveal melanomas were stained with the lectin Ulex europaeus agglutinin I (UEAI) and proliferating cell nuclear antigen (PCNA) to demonstrate simultaneously the tumor blood vessels and proliferating tumor cells. Indocarbocyanine (Cy3) was used as a fluorophore for UEAI and indodicarbocyanine (Cy5) was used for PCNA. Double labeled sections were examined with a laser scanning confocal microscope. Images of both stains were digitized at the same 5-microns intervals and each of the two images per interval was combined digitally to form one image. These combined images were visualized through voxel processing to study the relationship between melanoma cells expressing PCNA and various microcirculatory patterns. This technique produces images comparable to scanning electron microscopy of cast corrosion preparations while permitting simultaneous localization of melanoma cells expressing PCNA. The microcirculatory tree can be viewed from any perspective and the relationship between tumor cells and the tumor blood vessels can be studied concurrently in three dimensions. This technique is an alternative to cast corrosion preparations.

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